猪小肠黏膜下层与兔骨髓间充质干细胞的体外复合培养  

In vitro coculture of bone marrow mesenchymal stem cells with the small intestinal submucosa

作　　者：魏人前[1] 曹兴海[1] 邓睿[1] 杨志明[2] 谭波[2] 靳安民[3] 

机构地区：[1]佛山市第二人民医院骨科,广东省佛山市528322 [2]四川大学华西医院干细胞与组织工程研究室,四川省成都市610044 [3]南方医科大学珠江医院骨科,广东省广州市510000

出　　处：《中国组织工程研究》2012年第43期8083-8089,共7页Chinese Journal of Tissue Engineering Research

基　　金：广东省医学科研基金(广东省卫生厅)立项课题(A2010601);佛山市科技局立项项目(201008026)~~

摘　　要：背景:小肠黏膜下层细胞外基质材料免疫原性低,具有良好生物相容性,是构建单一结构工程化组织的较好支架材料。目的:观察体外兔骨髓间充质干细胞与猪小肠黏膜下层复合培养的生物相容性。方法:采用密度梯度离心法结合贴壁法分离纯化培养兔骨髓间充质干细胞,在其接种前用红色免疫荧光标记,然后将第2代已标记的兔骨髓间充质干细胞接种在猪小肠黏膜下层上。结果与结论:①组织学观察:骨髓间充质干细胞与小肠黏膜下层上复合培养1周时细胞呈单层生长,荧光显微镜下观察标记的骨髓间充质干细胞显示均匀红色荧光,复合培养2周细胞呈多层生长,并显示更密集的红色荧光。②扫描电镜观察:复合培养2d,骨髓间充质干细胞黏附于材料表面并伸展;复合培养1周,小肠黏膜下层被骨髓间充质干细胞分泌的胶原覆盖;2周后细胞在材料上已大量增殖形成融合,细胞连接紧密,细胞分泌大量基质,并分层。表明小肠黏膜下层与骨髓间充质干细胞具有良好的相容性。BACKGROUND： Small intestinal submucosa extracellular matrix is a kind of material used to build the single structural engineering scaffolds with low immune response and good biocompatibility. OBJECTIVE： To observe the biocompatibility of bone marrow mesenchymal stem cells growing on the small intestina submucosa in vitro. METHODS： The primary bone marrow mesenchymal stem cells were separated and purified by density gradient centrifugation combined with adherent methods. The cells were labeled using the red fluorescent cell linker kit before seeding. The passage 2 labeled bone marrow mesenchymal stem cells were seeded on the small intestinal submucosa.RESULTS AND CONCLUSION：Histological observation showed that the bone marrow mesenchymal stem cells formed a single layer on the surface of small intestinal submucosa after coculture for 1 week. The labeled bone marrow mesenchymal stem cells on the scaffold showed uniformly red fluorescence by fluorescence microscope. Two weeks later, the cells were in multi-layer, and displayed intensively red fluorescence. （2）Scanning electron microscope showed that, after 2 days of coculture, bone marrow mesenchymal stem cells were adherent to the small intestinal submucosa; 1 week later, the small intestinal submucosa was covered with the collagen secreted by bone marrow mesenchymal stem cells; 2 weeks later, the cells proliferated and fused on co-cultured small intestinal submucosa and closely connected, which showed stratifications and secreted a large number of matrix. The data obtained from the present study shows that bone marrow msenchymal stem cells and small intestinal submucosa have a good compatibility.

关 键 词：小肠黏膜下层 骨髓间充质干细胞 复合培养 组织工程 神经 标记 生物材料 生物相容性 

分 类 号：R318[医药卫生—生物医学工程]